A randomized controlled study of human serum albumin and serum substitute supplement as protein supplements for IVF culture and the effect on live birth rates

Meintjes, M.; Chantilis, S.J.; Ward, David C.; Douglas, James; Rodríguez, A.; Guerami, A.; Bookout, D.M.; Barnett, B.D.; Madden, James D.

doi:10.1093/humrep/den396

Cited by 60 publications

(36 citation statements)

References 18 publications

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“…Previous studies have demonstrated that both HSA and complex protein supplements are heterogeneous in nature [19] and thus represent variation that may impact MEA results. Specifically, complex proteins (SPS, LGPS) contain more transition metals than other protein supplements which may increase reactive-oxidant species and result in cell damage [18].…”

Section: Discussionmentioning

confidence: 99%

“…For human IVF, decreased oxygen concentration (5%) is both physiologic in the reproductive tract and associated with improved outcomes [13][14][15][16], yet many IVF clinics throughout the world still use atmospheric oxygen for culture and for QC testing [17]. The type of protein also varies in practice, thus, representing another variable that could influence assay sensitivity [18,19]. A third variation in embryo culture methods, group versus individual culture, influences QC sensitivity [10], and evidence suggests that group culture improves IVF outcomes, either through diffusion of unknown paracrine factors or maintenance of a stable microenvironment [20].…”

Section: Introductionmentioning

confidence: 99%

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Improved detection of mineral oil toxicity using an extended mouse embryo assay

Ainsworth

Fredrickson

Morbeck

2017

J Assist Reprod Genet

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Purpose Successful in vitro fertilization (IVF) relies on sound laboratory methods and culture conditions which depend on sensitive quality control (QC) testing. This study aimed to improve the sensitivity of mouse embryo assays (MEA) for detection of mineral oil toxicity. Methods Five experiments were conducted to study modifications of the standard mouse embryo assay (MEA) in order to improve sensitivity using clinical grade mineral oil with known peroxide concentrations. Assessment of blastocyst development at either 96 h or in an extended MEA (eMEA) to 144 h was tested in each experiment. In experiment 1, ability to detect peroxides in oil was compared in the MEA, eMEA, and cell number at 96 h. In experiment 2, serial dilutions of peroxide in oil were used along with time-lapse imaging to compare sensitivity of the morphokinetic MEA to the eMEA. Culture conditions that may affect assay sensitivity were assessed in experiments 3-5, which examined the effect of group versus individual culture, oxygen concentration, and protein supplementation. Results Extended MEA and cell counts identified toxicity not detected by the routine endpoint of blastocyst rate at 96 h. The eMEA was fourfold more sensitive than the standard MEA, and this sensitivity was similar to the morphokinetic MEA. Group culture had a protective effect against toxicity, while oxygen concentration did not affect blastocyst development. Protein supplementation with HSA had a protective effect on blastocyst development in eMEA.Conclusions The standard MEA used by manufacturers does not detect potentially lethal toxicity of peroxides in mineral oil. While group culture may mask toxicity, protein supplementation and oxygen concentration have minimal effect on assay sensitivity. The eMEA and time-lapse morphokinetic assessment are equally effective in detection of peroxide toxicity and thus provide manufacturers and end-users a simple process modification that can be readily adopted into an existing QC program.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Improved detection of mineral oil toxicity using an extended mouse embryo assay

Ainsworth

Fredrickson

Morbeck

2017

J Assist Reprod Genet

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“…In this study, we cultured hAECs with serum substitute supplement routinely used as culture media for protein supplements in gamete and embryo manipulation for assisted reproductive procedures (13).…”

Section: Introductionmentioning

confidence: 99%

Human Amniotic Epithelial Cells Cultured in Substitute Serum Medium Maintain Their Stem Cell Characteristics for Up to Four Passages

Evron

Goldman²,

Shalev

2011

Int J Stem Cells

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Background and Objectives: The common applied culture medium in which human amniotic epithelial cells (hAECs) maintain their stem cell characteristics contains fetal calf serum (FCS) and thus is not compatible with possible future clinical applications due to the danger of animal derived pathogens. To overcome this problem, we replaced FCS with serum substitute supplement, a serum substitute used in the in vitro fertilization for embryo development, in the common applied culture medium and cultured hAECs in this substitute serum medium (SSM). Methods and Results: Purity validation and characterization of freshly isolated and cultured hAECs was assessed through the expression of stem cell specific markers by RT-PCR (gene expression), by immunofluorescence staining and FACS (protein expression). Furthermore, karyotype was performed at passage four in order to exclude possible chromosome anomalies in hAECs cultured in SSM. The differentiation potential of hAECs into the cardiomyogenic lineage was tested through cardiac Troponin T expression by immunohistochemistry. hAECs cultured in SSM maintained expression of all the major pluripotent genes Sox-2, Oct-4 and Nanog as well as the expression of the embryonic stem cell specific surface antigens SSEA-4, SSEA-3 and TRA-1-60 over four passages. Using cardiac differentiation medium containing 10% serum substitute supplement, hAECs differentiated into cardiac troponin T expressing cells. Conclusions: We can conclude that, hAECs maintain their stem cell characteristics when cultured in SSM for up to 4 passages. This makes possible future clinical applications of these cells more feasible.

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“…Beckwith-Wiedemann syndrome or large-offspring syndrome known from veterinary embryo in vitro culture possibly resulting from imprinting disorders or potentially elongated culture of blastocysts, raises the question whether the culture media in human in vitro systems may alter the protein expression profile of developing embryos as well and therewith probably introducing a negative implanting capacity or even worse occurrence of syndromes in the offspring. Studies employing mouse and human embryos to investigate different culture media and their impact on embryo morphology, cleavage rates, blastocyst development and implantation rate revealed a considerable impact of the media and, respectively, of the resulting pregnancy rates [21,22,23,24]. Furthermore, since the introduction of the intracytoplasmic sperm injection technique in ART, the overall pregnancy rate in in vitro fertilization (IVF) cycles remained almost invariant within the last years indicating that yet another advancement in pregnancy rate might only be achieved by improving the embryo culture condition.…”

Section: Introductionmentioning

confidence: 99%

In vitro Culture Does Not Alter the Expression of Vascular Endothelial Growth Factor and Its Receptors in Single Murine Preimplantation Embryos

Dm¹,

Js²,

Sc³

et al. 2010

Gynecol Obstet Invest

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Background: In vitro culture of embryos, as widely used in assisted reproduction techniques, may influence embryonic development and subsequently the establishment of pregnancy. The aim of this study was to determine a potential influence of the in vitroculture regarding VEGF, VEGFR1 and VEGFR2 mRNA expression in developing single mouse embryos. Methods: Murine embryos were isolated on day 1 post coitus (p.c.) and cultivated for a developmental time course followed by examination for mRNA expression using RT-nested PCR. Furthermore, in vitro cultured blastocysts were compared to in vivo development at 101 h p.c. Results: At 101 h p.c. there were no significant differences between in vivo and in vitro cultured blastocysts regarding the expression of VEGF and its receptors. In the developmental time course, VEGF expression increased up to 94% in late blastocysts whereas the VEGF receptor expression remained low. Conclusions: This study showed that the in vitro culture did not alter the embryonic VEGF and VEGFR mRNA expression reassuring that the culture conditions in assisted reproduction techniques are well suited for maintaining the VEGF mRNA expression profile. Additionally, nearly 100% VEGF expression in late blastocysts highlights its importance for angiogenesis induction at the fetal-maternal interface.

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A randomized controlled study of human serum albumin and serum substitute supplement as protein supplements for IVF culture and the effect on live birth rates

Cited by 60 publications

References 18 publications

Improved detection of mineral oil toxicity using an extended mouse embryo assay

Improved detection of mineral oil toxicity using an extended mouse embryo assay

Human Amniotic Epithelial Cells Cultured in Substitute Serum Medium Maintain Their Stem Cell Characteristics for Up to Four Passages

In vitro Culture Does Not Alter the Expression of Vascular Endothelial Growth Factor and Its Receptors in Single Murine Preimplantation Embryos

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